樟芝之固態發酵培養及活性成分之研究

Studies on the Solid State Fermentation and Bioactive Components of Antrodia cinnamomea

劉艾淇(Ai-Chi Liu)

指導教授 ： 賴宏亮

國立屏東科技大學/農學院/農園生產系所/碩士(2014年)

https://doi.org/10.6346/NPUST.2014.00320

摘要

樟芝 (Antrodia cinnamomea) 為多孔菌科薄孔菌屬，只生長在臺灣特有種的牛樟樹 (Cinnamonum kanehirae Hay) 之中空內壁中，為一種木材褐腐菌。其成分包括多醣，三萜類化合物和苯環型化合物等，研究顯示，這些化合物具有抗氧化、抗腫瘤及免疫調節活性。由於其成分之藥理活性，目前已成為臺灣最昂貴的菇類。為了進一步提高生物活性之成分，本研究以ACS-1及ACE-1菌種，利用穀物作為固態培養基，進行固態發酵培養，探討兩種菌種於不同固態培養基及不同採收期間菌絲生長量及成分含量之變化。並以管柱層析方法及回收製備型HPLC (Recycling Preparative, HPLC) 進行樟芝成分分離與純化，並開發高效液相層析法 (High performance liquid chromatography, HPLC) 之成分分析法。結果顯示，ACS-1及ACE-1菌種皆於第三個月培養產量較高，其中ACS-1及ACE-1菌種皆以紅薏仁培養之菌絲體產量較高，分別為8.34±3.62 g/bottle D.W.及5.38±1.45 g/bottle D.W.。另外，由樟芝菌絲體乾燥分離純化得到Eburicoic acid、Dehydroeburicoic acid、15α-Acetyl-dehydrosulphurenic acid及Versisponic acid D，與4,7-Dimethoxy-5-methyl-1,3-benzodioxole (DMB) 及4-Acetylantroquinonol B (AQB) 六個成分做為指標成分進行成分定量分析，於不同培養時間之定量結果以第二個月各成分含量最高，DMB以糯米培養為15.96±1.74 mg/g D.W.；AQB以秈米培養為7.43±0.58 mg/g D.W.；Eburicoic acid以臺梗九號培養為9.39±1.08 mg/g D.W.；Dehydroeburicoic acid以紅薏仁培養為6.76±1.16 mg/g D.W.；15α-Acetyl-dehydrosulphurenic acid以臺梗九號培養為5.21±0.17 mg/g D.W.；Versisponic acid D以臺梗九號培養為0.99±0.02 mg/g D.W.之含量為最高。整體而言，不同培養基培養之樟芝菌絲體的成分含量各不相同，以糯米做為固態培養基培養樟芝菌絲體之各成分含量加總為最高，本論文實驗結果可供利用穀物固態培養基培養樟芝菌絲體之生產作為參考，以提高菌絲體產量及各活性成分之含量，建立優良產物品質及提供開發保健食品或醫藥之參考。
關鍵字: 樟芝、活性成分、管柱層析、高效液相層析法

關鍵字

樟芝 ； 活性成分 ； 管柱層析 ； 高效液相層析法

並列摘要

Antrodia cinnamomea, an important endemic species in Taiwan, growth only in the inner cavity of the rotten trunk of Cinnamonum kanehirae Hay. As a wood brown rot fungi. Main A. cinnamomea ingredients are polysaccharides, triterpenoids, and Benzenoids. Studies indicated that these compounds have antioxidant, antitumor, and immunomodulatory activities. Due to the pharmacological activities, A. cinnamomea has become the most expensive mushroom in Taiwan. To further improve the yields of the active compounds and biological activities, two strain of A. cinnamomea, ACS-1 and ACE-1, were used and components of their mycelia grown on different culturing medium at different harvest time were investigated using column chromatography and high performance liquid chromatography (HPLC). The components were separated and purified and quantitative ingredients analysis were conducted. The results showed that ACS-1 and ACE-1 strains had higher yields of the compounds listed above when were cultured for three months. The highest yields of ACS-1 and ACE-1mycelia grown on red coix were of 8.34 ± 3.62 g/bottle D.W. and 5.38 ± 1.45 g/bottle D.W., respectively. Meanwhile, Eburicoic acid, Dehydroeburicoic acid, 15α-Acetyl-dehydrosulphurenic acid, Versisponic acid D, 4,7-Dimethoxy-5-methyl-1,3-benzodioxole (DMB), and 4-Acetylantroquinonol B (AQB) were used as indicators to process separation and purification on dry mycelia. Results showed that highest content of each component was found at the culturing time of the second month. Moreover, mycelia grown on Tai-sen glutinous 2 had the highest content of DMB (15.96 ± 1.74 mg/g D.W.). Meanwhile, the highest compound yields in dry A. cinnamomea mycelia and respective culturing medium were listed as follow: Taichung sen 10: AQB of 7.43± 0.58 mg/g D.W.; Taiken 9: Eburicoic acid of 9.39 ± 1.08 mg/g D.W.; red coix: Dehydroeburicoic acid of 6.76 ± 1.16 mg/g D.W.; Taiken 9: 15 α-Acetyl-dehydrosulphurenic acid of 5.21 ± 0.17 mg/g D.W.; Taiken 9: Versisponic acid D of 0.99 ± 0.02 mg/g D.W. Totally, compounds in A. cinnamomea mycelia cultivated on different media were varied. The mycelia grown on Tai-sen glutionous 2, as a solid medium, had the highest total compound yields. Using cereal solid substrates, the results of this study can be used as a reference for culturing A. cinnamomea mycelia to have higher yields and bioactivities and a reference for the developing health food or medicine.

Key words：Antrodia cinnamomea, bioactive components, column
chromatography, high performance liquid chromatography

並列關鍵字

Antrodia cinnamomea ； bioactive components ； column chromatography ； high performance liquid chromatography

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